MSC/ADSC Isolation From Human Lipoaspirate
(Miltenyi Biotec)This protocol describes the preparation of MSCs from human thigh or abdomen lipoaspirate obtained from cosmetic surgery. Briefly, the lipoaspirate is first washed thoroughly in phosphate-buffered saline (PBS) before being subjected to enzymatic digestion using collagenase in order to obtain a single-cell suspension. After digestion, the centrifuged cell pellet, termed the stromal vascular fraction (SVF), is resuspended in NH Expansion Medium before serial filtration through 100 μm and then 40 μm nylon filters. The content of mononuclear cells is then counted and the cell sample taken into culture in NH Expansion Medium. MSCs adhere to plastic surfaces within 24 hours after which the medium should be exchanged with fresh NH Expansion Medium.http://www.miltenyibiotec.com/download/protocols_c...Added: Wed Jan 28 2009, Hits: 2958, Reviews: 0Write review